Glechoma longituba volatile oil as well as preparation method and application thereof

Through tissue culture and Na2SO4 stress treatment, four new volatile oils were prepared, which solved the problem of unstable volatile oil components, improved its antioxidant and lowered blood lipids, and protected the function of vascular endothelial cells.

CN120399803APending Publication Date: 2025-08-01HUBEI UNIV FOR NATITIES
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Patent Information

Application Number
CN202510681421.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-05-26
Publication Date
2025-08-01

AI Technical Summary

Technical Problem

The composition and quality of volatile oil of volatile oils of lianqiancao are greatly affected by factors such as plant growth season, soil, climate, altitude, pests, pesticides and storage, resulting in unstable medicinal effects. It is difficult for the existing technology to effectively control and improve its antioxidant, lower blood lipids and protect vascular endothelial cells.

Method used

Four new volatile oils were prepared by tissue culture, liquid culture and Na2SO4 stress treatment. The specific steps include transferring the sterile seedlings of lianqian tissue culture into Japanese garden general nutrient solution for cultivation, cutting them into small sections and inserting them into hydroponics, and steam distillation is performed after Na2SO4 stress to extract the volatile oil.

Benefits of technology

It significantly increases the content of specific compounds in volatile oils, improves its antioxidant ability and therapeutic effect on cardiovascular and cerebrovascular diseases, especially the function of lowering blood lipids and protecting vascular endothelial cells.

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Abstract

The invention relates to four novel glechoma longituba volatile oils and a preparation method thereof, and belongs to the technical field of biology. Transferring tissue-cultured glechoma longituba aseptic seedlings into a 1 / 2 Japanese garden test nutrient solution for culturing to obtain hydroponic glechoma longituba; the method comprises the following steps: carrying out stress water culture on glechoma longituba for 72 hours by using 0.4 wt / v% of Na2SO4, carrying out stress water culture on glechoma longituba for 4 hours, 48 hours and 96 hours by using 1.6 wt / v% of Na2SO4, harvesting stems and leaves of glechoma longituba, and extracting volatile oil by using a steam distillation method to obtain four types of volatile oil in total. The glechoma longituba volatile oil obtained by the invention is used for preparing medicines for resisting oxidation, reducing blood fat, protecting vascular endothelial cells, resisting cardiovascular and cerebrovascular system related diseases such as atherosclerosis and the like.
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Description

Technical Field

[0001] The present invention relates to four novel volatile oils from Herba Lycopodii and methods for preparing them, belonging to the field of biotechnology. Herba Lycopodii is subjected to tissue culture followed by liquid culture, and the stems and leaves of the Herba Lycopodii are subjected to Na2SO4 stress treatment. The volatile oils are extracted by steam distillation to prepare four volatile oils having compositions different from those of unstressed Herba Lycopodii. The Herba Lycopodii volatile oils obtained by the present invention are used to prepare drugs for lowering blood lipids, protecting vascular endothelial cells, and combating cardiovascular and cerebrovascular diseases such as atherosclerosis. Background Art

[0002] Lianqiancao is a plant of the genus Huoxuedan in the family Lamiaceae. Glechoma longituba (Nakai) Kupr.] Dried aerial parts, sun-dried, or used fresh. Herba Lycopodii has a cool, pungent, slightly bitter, and slightly cold nature. It enters the liver, kidney, and bladder meridians. Herba Lycopodii has choleretic, diuretic, antibacterial, anti-inflammatory, hypoglycemic, anti-tumor, and anti-diarrheal effects. It is primarily used to treat icteric hepatitis, cholecystitis, cholelithiasis, kidney stones, bladder stones, and mumps. Herba Lycopodii volatile oil is one of its main medicinal components. However, as a secondary metabolite, volatile oil is susceptible to factors such as the plant's growing season, soil, climate, altitude, pests and diseases, pesticides, and the harvesting and storage of medicinal materials. Its composition and quality vary widely and are uncontrollable. Salt stress can increase the content of active ingredients in medicinal plants, but its effects on the composition and function of volatile oils remain unknown. The present invention solves the technical difficulties in the preparation of volatile oil from Scutellaria baicalensis through tissue culture, liquid culture and salt stress, and obtains four new volatile oils from Scutellaria baicalensis. The volatile oils from Scutellaria baicalensis obtained by the invention are used to prepare drugs for cardiovascular and cerebrovascular system-related diseases such as antioxidant, lipid-lowering, endothelial cell protection and anti-atherosclerosis. Summary of the Invention

[0003] One of the purposes of the present invention is to provide a method for preparing Herba Lycopodii volatile oil-1 through tissue culture, liquid culture and Na2SO4 stress.

[0004] The Glechoma longituba essential oil-1 described in the present invention has 15 new compounds among the top 50 compounds in terms of content, compared with the Glechoma longituba essential oil without stress. The 15 new compounds are 1,3,4-trimethyl-3-cyclohexene-1-carbaldehyde (CAS No.: 040702-26-9), Z,Z,Z-1,5,9,9-tetramethyl-1,4,7-cycloundecatriene (CAS No.: 1000062-61-9), β-ocimene (CAS No.: 013877-91-3), hexahydro-2,5-methano-1H-indene-7(4H)-one (CAS No.: 027567-85-7), ethyl decanoate (CAS No.: 000110-38-3), nonenal (000124-19-6), 2-methyl-3-phenylpropenal (CAS No.: 000101-39-3), 2-methyloctane (CAS No.: 003221-61-2), trans-2-methyl-5-(1-methylethenyl)cyclohexanone (CAS No.: 005948-04-9), 1-vinyl-1-methyl-2,4-bis(1-methylethenyl)cyclohexane (CAS No.: 110823-68-2), 7,11-dimethyl-3-methylene-1,6,10-dodecatriene (CAS No.: 077129-48-7), (1-α,2-β,4-β)-1-vinyl-1-methyl-2,4-bis(1-methylethenyl)cyclohexene (CAS No.: 033880-83-0), (Z,Z)-α-farnesene (1000293-03-1), 2-methyl-5-(1-methylethyl)phenol (000499-75-2), cis-p-menth-2,8-dien-1-ol (003886-78-0); there are 6 compounds with significantly increased content ( P <0.05), which are as follows: caryophyllene (CAS No.: 000087-44-5), (E)-β-farnesene (CAS No.: 018794-84-8), [S-(R,S -3-(1,5-dimethyl-4-hexenyl)-6-methylenecyclohexene (CAS No.: 020307-83-9), 1-(1,5-dimethyl-4-hexenyl)-4-methylbenzene (CAS No.: 000644-30-4), [S-(R,S )]-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS No.: 000495-60-3), (1S-cis)-1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-naphthalene (CAS No.: 000483-76-1).

[0005] The method for obtaining Glechoma longituba essential oil-1 is to transfer aseptic tissue-cultured seedlings of Glechoma longituba into 1 / 2 strength Japanese standard nutrient solution for cultivation; then cut the Glechoma longituba seedlings into small segments and insert them into 1 / 2 strength Japanese standard nutrient solution for cultivation until more than 15 nodes at the stem tip are obtained to get hydroponic Glechoma longituba. After subjecting the hydroponic Glechoma longituba to Na2SO4 stress, essential oil is extracted by steam distillation method.

[0006] In the said method, the liquid culture temperature is 20-30 °C.

[0007] In the said method, the liquid culture time is 25-60 d.

[0008] In the said method, the concentration of Na2SO4 is 1.2 wt / v% - 2.0 wt / v%.

[0009] In the said method, the time of Na2SO4 stress is 2-6 h.

[0010] The second object of the present invention is to provide a method for preparing Glechoma longituba essential oil-2 through tissue culture, liquid culture and Na2SO4 stress.

[0011] For the Glechoma longituba essential oil-2 of the present invention, among the top 50 compounds in terms of content, compared with the essential oil of Glechoma longituba without stress, 10 new compounds are added, namely cis-p-mentha-2,8-dien-1-ol (CAS number: 003886-78-0), β-pinene (CAS number: 000123-35-3), β-ocimene (CAS number: 013877-91-3), ethyl caprate (CAS number: 000110-38-3), 1,7,7-trimethyl-bicyclo[2.2.1]hept-2-ene (CAS number: 000464-17-5), (Z,Z)-α-farnesene (CAS number: 1000293-03-1), (6R,7aR)-3,6-dimethyl-5,6,7,7a-tetrahydrobenzofuran-2(4H)-one (CAS number: 038049-04-6), 1-ethoxy-4-ethylbenzene (CAS number: 001585-06-4), 2-methyl-5-(1-methylethyl)phenol (CAS number: 000499-75-2), 1-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2-buten-1-one (CAS number: 035044-68-9); the content increases significantly ( PThere are 6 compounds with a content of <0.05), namely: linalool (CAS No.: 000078-70-6), 3-methyl-6-(1-methylethenyl)-cyclohexen-1-one (CAS No.: 000491-09-8), 10,10-dimethyl-2,6-dienylbicyclo[7.2.0]undecane (CAS No.: 357414-37-0), [S-(R,S )-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS No.: 000495-60-3), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS No.: 007216-56-0), (1S-cis)-1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-naphthalene (CAS No.: 000483-76-1).

[0012] The method for obtaining Glechoma longituba volatile oil-2 is to transfer the aseptic seedlings of Glechoma longituba tissue culture into 1 / 2 Japanese Horticultural Test Commodity Nutrient Solution for cultivation; then cut the Glechoma longituba seedlings into small segments and insert them into 1 / 2 Japanese Horticultural Test Commodity Nutrient Solution for cultivation until more than 15 nodes at the stem tip are obtained to get hydroponic Glechoma longituba. After subjecting the hydroponic Glechoma longituba to Na2SO4 stress, the volatile oil is extracted by steam distillation method.

[0013] In the said method, the liquid culture temperature is 20 - 30 °C.

[0014] In the said method, the liquid culture time is 25 - 60 d.

[0015] In the said method, the concentration of Na2SO4 is 0.2 wt / v% - 0.6 wt / v%.

[0016] In the said method, the time of Na2SO4 stress is 68 - 76 h.

[0017] The third object of the present invention is to provide a method for preparing Glechoma longituba volatile oil-3 through tissue culture, liquid culture and Na2SO4 stress.

[0018] The Glechoma longituba volatile oil-3 of the present invention has 11 new compounds among the top 50 compounds in terms of content, compared with the Glechoma longituba volatile oil without stress, namely: (1S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene (CAS No.: 007785-26-4), β-ocimene (CAS No.: 013877-91-3), ethyl caprate (CAS No.: 000110-38-3), 2-methyl-3-phenyl-2-propenal (CAS No.: 000101-39-3), aromadendrene (CAS No.: 000489-39-4), α-terpineol (CAS No.: 000098-55-5), cis-3-hexenyl cis-3-hexenoate (CAS No.: 061444-38-0), (Z)-2-(3,3-dimethylcyclohexenyl)-ethanol (CAS No.: 026532-23-0), neodihydrocarveol (CAS No.: 018675-34-8), 1-methyl-3-(1-methylethenyl)cyclohexene (CAS No.: 000499-03-6), trans-2-methyl-5-(1-methylethenyl)cyclohexanone (CAS No.: 005948-04-9); the contents of the following 4 compounds are significantly increased ( P <0.05): D-limonene (CAS No.: 005989-27-5), trans-5-methyl-2-(1-methylethenyl)-cyclohexanone (CAS No.: 029606-79-9), caryophyllene (CAS No.: 000087-44-5), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS No.: 007216-56-0).

[0019] The method for obtaining Glechoma longituba volatile oil-3 is to transfer the aseptic seedlings of Glechoma longituba tissue culture into 1 / 2 Japanese Horticultural Standard Solution for culture; then cut the Glechoma longituba seedlings into small segments and insert them into 1 / 2 Japanese Horticultural Standard Solution for culture until more than 15 nodes at the stem tip are obtained to get hydroponic Glechoma longituba. After subjecting the hydroponic Glechoma longituba to Na2SO4 stress, the volatile oil is extracted by steam distillation method.

[0020] In the said method, the liquid culture temperature is 20-30 °C.

[0021] In the said method, the liquid culture time is 25-60 d.

[0022] In the said method, the concentration of Na2SO4 is 1.2 wt / v%-2.0 wt / v%.

[0023] In the said method, the stress time of Na2SO4 is 92-100 h.

[0024] A fourth object of the present invention is to provide a method for preparing Glechoma longituba essential oil-4 through tissue culture, liquid culture, and Na2SO4 stress.

[0025] Among the top 50 compounds in terms of content, compared with the essential oil of Glechoma longituba without stress, 11 new compounds are added to the Glechoma longituba essential oil-4 of the present invention, namely: β-ocimene (CAS number: 013877-91-3), ethyl caprate (CAS number: 000110-38-3), Z,Z,Z-1,5,9,9-tetramethyl-1,4,7-cycloundecatriene (CAS number: 1000062-61-9), hexahydro-2,5-methano-1H-indene-7(4H)-one (CAS number: 027567-85-7), 2-isopropyl-5-methyl-3-cyclohexen-1-one (CAS number: 1000155-47-0), (+)-4-carene (CAS number: 029050-33-7), 3-methyl-6-(1-methylethenyl)-cyclohexene (CAS number: 000586-63-0), 7,11-dimethyl-3-methylene-1,6,10-dodecatriene (CAS number: 077129-48-7), 1,5,5-trimethyl-6-methylene-cyclohexene (000514-95-4), 5-methyl-3-(1-methylethenyl)cyclohexene (CAS number: 056816-08-1), cis-3-hexenyl cis-3-hexenoate (CAS number: 061444-38-0). There are 5 compounds with significantly increased content ( P <0.05), namely: trans-5-methyl-2-(1-methylethenyl)-cyclohexanone (CAS number: 029606-79-9), myrcene (CAS number: 000087-44-5), linalool (CAS number: 000078-70-6), [S-(R,S )]-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS number: 000495-60-3), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS number: 007216-56-0).

[0026] The method for obtaining Glechoma longituba essential oil-4 is to transfer the aseptic seedlings of Glechoma longituba in tissue culture into 1 / 2 Japanese Horticultural Test Commodity Solution for culture; then cut the Glechoma longituba seedlings into small segments and insert them into 1 / 2 Japanese Horticultural Test Commodity Solution for culture until more than 15 nodes at the stem tip are obtained to get hydroponic Glechoma longituba. After subjecting the hydroponic Glechoma longituba to Na2SO4 stress, the essential oil is extracted by steam distillation.

[0027] In the said method, the temperature for liquid culture is 20-30 °C.

[0028] In the method described above, the liquid culture time is 25 - 60 d.

[0029] In the method described above, the concentration of Na2SO4 is 1.2 wt / v% - 2.0 wt / v%.

[0030] In the method described above, the time of Na2SO4 stress is 44 - 52 h. Description of the Drawings

[0031] Figure 1 It is the GC - MS spectrum of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO). A represents non - stressed, B represents stressed with 0.4 wt / v% Na2SO4, C represents stressed with 0.8 wt / v% Na2SO4, D represents stressed with 1.2 wt / v% Na2SO4, and E represents stressed with 1.6 wt / v% Na2SO4.

[0032] Figure 2 It is the DPPH free radical scavenging ability of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO). A represents GA, and B represents GLVO.

[0033] Figure 3 It is the ABTS free radical scavenging ability of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO). A represents GA, and B represents GLVO.

[0034] Figure 4 It is the FRAP value of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO). A represents GA, and B represents GLVO.

[0035] Figure 5 It is the total reducing power of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO). A represents GA, and B represents GLVO.

[0036] Figure 6 It is the effect of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO) on reducing the total cholesterol (TC) in mice fed with high - fat diet. A represents the blank group (GLVO - 1), B represents volatile oil - 1 (GLVO - 2), and C represents volatile oil - 2 (GLVO - 3).

[0037] Figure 7 It is the effect of the volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO) on reducing the triacylglycerol (TG) in mice fed with high - fat diet. A represents the blank group (GLVO - 1), B represents volatile oil - 1 (GLVO - 2), and C represents volatile oil - 2 (GLVO - 3).

[0038] Figure 8To study the proliferative effect of Glechoma longituba volatile oil (GLVO) on vascular endothelial cells, Group A was the blank group (GLVO-1), Group B was volatile oil-1 (GLVO-2), Group C was volatile oil-2 (GLVO-3), Group D was volatile oil-3 (GLVO-4), and Group E was volatile oil-4 (GLVO-5).

[0039] Figure 9 To study the repair effect of Glechoma longituba volatile oil (GLVO) on vascular endothelial cells damaged by LPS, Group A was the blank group (GLVO-1), volatile oil-2 (GLVO-3) and volatile oil-4 (GLVO-5). Group B was volatile oil-1 (GLVO-2) and volatile oil-3 (GLVO-4).

[0040] Figure 10 To study the effect of Glechoma longituba volatile oil (GLVO) in reducing the MDA level in vascular endothelial cells damaged by LPS, Group A was the blank group (GLVO-1), volatile oil-2 (GLVO-3) and volatile oil-4 (GLVO-5). Group B was volatile oil-1 (GLVO-2) and volatile oil-3 (GLVO-4). Specific Embodiments

[0041] The present invention will be further described below in conjunction with embodiments. The advantages and features of the present invention will become clearer as the description proceeds. However, these examples are merely exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that modifications or substitutions to the details and forms of the technical solutions of the present invention without departing from the spirit and scope of the present invention all fall within the protection scope of the present invention. Chemical reagents and the like used in the specification and embodiments are operated under conventional experimental conditions or according to the instructions provided by the suppliers unless otherwise specified.

[0042] Example 1: Preparation of Glechoma longituba Hydroponic Seedlings Take Glechoma longituba tissue culture seedlings inoculated for 25 days and with 5 - 8 leaves grown. Rinse the medium on the roots with sterile water, transfer them to 1 / 2 Japanese Horticultural Test nutrient solution, and culture at 25°C for 60 days until there are more than 15 nodes at the stem tip. Cut the stem nodes and culture them by cuttage in 1 / 2 Japanese Horticultural Test nutrient solution for hydroponics. The hydroponic method is the static floating plate hydroponic method. The specific operation is as follows: Wash the Glechoma longituba stem segments about 3 - 5 cm long with water, gently pass them through the plastic plate with pre-punched holes, immerse the lower end of the stem segments in the culture solution, and culture at 25°C. Replace the nutrient solution every 7 days in the early stage of culture and every 3 days in the later stage. When replacing the nutrient solution, rinse the roots of the plants with clean water and clean the rotten leaves. Regularly supplement the lost water or nutrient solution, observe and record the growth status of the plants every 2 days, and culture for a total of 28 days. This is the Glechoma longituba hydroponic seedling of the present invention.

[0043] Example 2: Salt Stress and Volatile Oil Extraction of Hydroponic Glechoma longituba Seedlings When the hydroponic Glechoma longituba seedlings are cultured to more than 15 nodes, different stress treatments are carried out. The concentrations of anhydrous Na2SO4 for stress treatment are: 0.4 wt / v%, 0.8 wt / v%, 1.2 wt / v% and 1.6 wt / v% respectively. The treatment without adding anhydrous Na2CO3 is the control group (i.e., CK group), and the control group is continuously cultured with nutrient solution. After stress for 4, 8, 12, 24, 48, 72 and 96 h respectively, the stems and leaves of Glechoma longituba are harvested, and the volatile oil is extracted by steam distillation method.

[0044] Example 3: Analysis of Volatile Oil Components of Glechoma longituba The HS-SPME-GC-MS technology is used to determine the components of the volatile oil of Glechoma longituba.

[0045] The volatile oil of Glechoma longituba is diluted 600 times with n-hexane. Take 10 μL of the volatile oil solution and add it into a headspace screw-cap sample bottle, place it in a 40 °C water bath for 10 min to equilibrate, and then push out the fiber head for extraction for 30 min. Excess water is removed with anhydrous Na2SO4 during the extraction process.

[0046] A non-polar capillary column HP-5 (0.32 mm × 30 m, 0.25 μm) is used; the inlet temperature is 250 °C; helium is selected as the carrier gas with a flow rate of 1 mL / min; the programmed temperature rise (initial temperature 60 °C, hold for 2 min, temperature rise rate 5 °C / min, rise to 220 °C, hold for 3 min); splitless injection; the scanning mode is full scan.

[0047] The MS conditions are that the mass spectrometry interface temperature is 250 °C, the EI ionization mode is adopted, the ion source temperature is 230 °C, the electron energy is 70 eV, the quadrupole temperature is 150 °C, the scanning range is 30 - 450 amu (mass-to-charge ratio), and the solvent delay is 7 min. The chemical components in the volatile oil of Glechoma longituba are qualitatively analyzed through NIST14.L spectral library matching.

[0048] R language is used to perform multivariate statistical analysis on the volatile oil components of Glechoma longituba, and the comprehensive medicinal potential score of the volatile oil adopts the principal component factor analysis method.

[0049] In Figure 1 Through GC-MS analysis, compared with the control group (i.e., the non-stressed group), after Na2SO4 stress, the components of the volatile oil of Glechoma longituba have changed. A, B, C, D, and E represent the control group, 0.4 wt / v%, 0.8 wt / v%, 1.2 wt / v% and 1.6 wt / v% Na2SO4 stress groups respectively.

[0050] Table 1 shows the results of the top 10 rankings of the comprehensive medicinal potential scores of volatile oils. It can be seen from the table that the four volatile oils with the highest comprehensive medicinal potential scores are those under 1.6 wt / v% stress for 4 h, 0.4 wt / v% stress for 72 h, 1.6 wt / v% stress for 96 h, and 1.6 wt / v% stress for 48 h.

[0051] Table 1 Comprehensive scores of the top 10 rankings of the medicinal potential of volatile oils Group Comprehensive Score Rank 1.6%_4h 2.84505116 1 0.4%_72h 1.32386621 2 1.6%_96h 1.1261643 3 1.6%_8h 1.1229352 4 1.6%_48h 1.0089238 5 0.8%_96h 0.86643131 6 0.4%_24h 0.75167756 7 0.4%_96h 0.60052122 8 CK 0.58861802 9 1.2%_96h 0.52735035 10 Tables 2 - 5 show the top 50 compounds ranked by the component contents of Glechoma longituba volatile oil - 1, Glechoma longituba volatile oil - 2, Glechoma longituba volatile oil - 3, and Glechoma longituba volatile oil - 4 respectively. The ↑ and ↓ in the table indicate that the compound contents are significantly increased or decreased ( P <0.05).

[0052] Table 2 The top 50 compounds with the highest contents in Glechoma longituba volatile oil - 1 Table 3 The top 50 compounds with the highest contents in Glechoma longituba volatile oil - 2 Table 4 The top 50 compounds with the highest contents in Glechoma longituba volatile oil - 3 Table 5 The top 50 compounds with the highest contents in Glechoma longituba volatile oil - 4 Example 3: Detection of the in vitro antioxidant activity of Glechoma longituba volatile oil 1 Determination of the DPPH free radical scavenging ability Mix 100 μL of the sample solution with 100 μL of the DPPH solution (0.1 mmol / L), react in the dark at room temperature for 30 min, measure the absorbance at 517 nm, and record it as As; the absorbance after mixing 100 μL of the sample solution with 100 μL of absolute ethanol is Ac; the absorbance after mixing 100 μL of the solvent with 100 μL of the DPPH solution is Ao. Use gallic acid (GA) as the standard substance. The DPPH free radical scavenging rate is calculated according to the following formula: DPPH free radical scavenging rate (%) = [1 - (As - Ac) / A0] × 100 The results are shown in Figure 2, in the range of 0.166 - 10.625 μmol / L of GA, the DPPH radical scavenging rate showed a linear correlation with the concentration. For GLVO - 1 (control group), in the range of 0.3125 - 20 mg / mL, the DPPH radical scavenging rate showed a linear correlation with the concentration; for GLVO - 2 (1.6 wt / v% stress for 4 h, volatile oil - 1), in the range of 0.0781 - 2.5 mg / mL, the DPPH radical scavenging rate showed a linear correlation with the concentration; for GLVO - 3 (0.4 wt / v% stress for 72 h, volatile oil - 2), in the range of 0.3125 - 20 mg / mL, the DPPH radical scavenging rate showed a linear correlation with the concentration. The half - clearing concentration CC 50 of GLVO - 1 (control group) was 13.10 mg / mL, and the CC 50 of GLVO - 2 (1.6 wt / v% stress for 4 h, volatile oil - 1) was 1.18 mg / mL, and the half - clearing concentration CC 50 of GLVO - 3 (0.4 wt / v% stress for 72 h, volatile oil - 2) was 11.88 mg / mL.

[0053] 2 Determination of ABTS radical scavenging ability Mix 140 mmol / L potassium persulfate solution with 7 mmol / L ABTS solution and react for 12 - 16 h under dark conditions at room temperature. Before use, dilute the mixed solution with absolute ethanol to an absorbance value of 0.7 ± 0.02 at 734 nm. Mix 50 μL of the sample solution with 150 μL of ABTS• + solution, react for 10 min at room temperature, and measure the absorbance value at 734 nm, denoted as As; the absorbance value after mixing 50 μL of the solvent with 150 μL of ABTS• + solution is Ao. Using GA as the standard substance, draw a standard curve. The ABTS radical scavenging rate is calculated according to the following formula: ABTS radical scavenging rate (%) = [1 - (A0 - As) / A0]×100 The results are as Figure 3As shown, in the range of 0.664 - 42.5 μmol / L of GA, the ABTS radical scavenging rate is linearly correlated with the concentration. For GLVO-1 (control group), in the range of 0.625 - 20 mg / mL, the ABTS radical scavenging rate is linearly correlated with the concentration; for GLVO-2 (0.4 wt / v% stress for 72 h, volatile oil - 2), in the range of 0.0781 - 2.5 mg / mL, the ABTS radical scavenging rate is linearly correlated with the concentration; for GLVO-3 (0.4 wt / v% stress for 72 h, volatile oil - 2), in the range of 0.3125 - 10 mg / mL, the ABTS radical scavenging rate is linearly correlated with the concentration. The half - clearing concentration CC 50 of GLVO-1 (control group) is 10.61 mg / mL, and the CC 50 of GLVO-2 (1.6 wt / v% stress for 4 h, volatile oil - 1) is 1.39 mg / mL, and the half - clearing concentration CC 50 of GLVO-3 (0.4 wt / v% stress for 72 h, volatile oil - 2) is 4.81 mg / mL.

[0054] 3 Determination of Ferric ion reducing antioxidant power (FRAP) An acetate buffer solution (300 mmol / L, pH 3.6), a solution of TPTZ (10 mmol / L) in HCl (40 mmol / L) and a FeCl3 solution (20 mmol / L) were mixed in a ratio of 10:1:1. 40 μL of the sample solution was mixed with 160 μL of the FRAP reagent, reacted at 37 °C for 30 min, and the absorbance was measured at 593 nm. Using GA as the standard substance, a standard curve was plotted.

[0055] The results are as Figure 4 shown. In the range of 0.166 - 85 μmol / L of GA, the FRAP value is linearly correlated with the concentration, with R 2 = 0.9989, and the linear equation is y = 9.941x + 0.103. For GLVO-1, in the range of 0.3125 - 10 mg / mL, the FRAP value is linearly correlated with the concentration, with R 2 = 0.9818, and the linear equation is y = 0.052x + 0.116. For GLVO-2, in the range of 0.3125 - 10 mg / mL, the FRAP value is linearly correlated with the concentration, with R 2= 0.989, and the linear equation is y = 0.1935x + 0.2431. In the range of 0.3125 - 10 mg / mL for GLVO-3, the FRAP value is linearly correlated with the concentration, and R 2 = 0.9823, and the linear equation is y = 0.083x + 0.112. As can be seen from the figure, the FRAP of GLVO-2 and GLVO-3 is higher than that of GLVO-1.

[0056] 4 Determination of total reducing power Mix 0.5 mL of the sample solution successively with 0.5 mL of PBS buffer solution and K3Fe(CN)6 solution, react at 50 °C in a water bath for 20 min. After the reaction, cool to room temperature and then add 0.5 mL of TCA solution, centrifuge (3000 r / min, 10 min), take 0.5 mL of the supernatant and mix it with 0.5 mL of distilled water and 0.1 mL of FeCl3 solution. After standing for 10 min, measure the absorbance at 700 nm. Use GA as the standard substance.

[0057] The results are as Figure 5 shown. In the range of 1.328 - 170 μmol / L for GA, the total reducing power is linearly correlated with the concentration, and R 2 = 0.9966, and the linear equation is y = 4.124x + 0.065. In the range of 0.3125 - 20 mg / mL for GLVO-1, the total reducing power is linearly correlated with the concentration, and R 2 = 0.9828, and the linear equation is y = 0.020x + 0.105. In the range of 0.3125 - 20 mg / mL for GLVO-2, the total reducing power is linearly correlated with the concentration, and R 2 = 0.9932, and the linear equation is y = 0.032x + 0.1138. In the range of 0.3125 - 20 mg / mL for GLVO-3, the total reducing power is linearly correlated with the concentration, and R 2 = 0.9935, and the linear equation is y = 0.023x + 0.089. The total reducing power of GLVO-2 is higher than that of GLVO-1, and the total reducing power of GLVO-3 is slightly higher than that of GLVO-1.

[0058] Example 4: Hypolipidemic effect of volatile oil from Glechoma longituba (Nakai) Kupr. Five-week-old male C57BL / 6 mice in good growth condition were selected and adaptively fed in the laboratory for 1 week. The mice were randomly divided into 13 groups: CK group (intragastric administration with normal diet + 0.5% sodium carboxymethylcellulose solution), model group (intragastric administration with high-fat diet + 0.5% sodium carboxymethylcellulose solution), positive control group 1 (intragastric administration with high-fat diet + 3 mg / kg / d lovastatin), positive control group 2 (intragastric administration with high-fat diet + 500 mg / kg / d VE), low-dose group (intragastric administration with high-fat diet + 50 mg / kg / d GLVO, with one group each for GLVO-1, GLVO-2, and GLVO-3), medium-dose group (intragastric administration with high-fat diet + 100 mg / kg / d GLVO, with one group each for GLVO-1, GLVO-2, and GLVO-3), high-dose group (intragastric administration with high-fat diet + 200 mg / kg / d GLVO, with one group each for GLVO-1, GLVO-2, and GLVO-3), with 10 mice in each group. GLVO-1, GLVO-2, and GLVO-3 represent the control group, 1.6 wt / v% stress for 4 h (volatile oil-1), and 0.4 wt / v% stress for 72 h (volatile oil-2) respectively. They were housed in individual ventilated cages using single-cage housing, with an air flow rate of 0.05 - 0.13 m / s, an air change rate of 20 - 70 N / h, a temperature of 20 - 22 °C, natural lighting, and unrestricted drinking water. Starting from the 4th week of administration, blood was collected from the orbital venous plexus of the mice once a week. The mice were fasted for 8 h before blood collection, with free access to drinking water. After blood collection, the blood was placed on ice for 25 min and then centrifuged (3500 r / min, 15 min), and the supernatant was taken to detect the contents of total cholesterol (TC) and triglyceride (TG). The administration lasted for 12 weeks in total.

[0059] The effect of volatile oil of Glechoma longituba (Nakai) Kupr. (GLVO) on TC in mice fed with high-fat diet is as Figure 6 shown. In the first 7 weeks, the TC levels in the model group and the administration groups gradually increased and were significantly higher than those in the CK group ( P< P < 0.05). After the 7th week, the TC levels in the mice of each administration group began to decrease and were significantly lower than those in the model group starting from the 8th week ( P< P < 0.05), but were still significantly higher than those in the CK group ( P< P < 0.05). When the treatment duration reached 10 weeks, the TC levels in each administration group had no significant difference from those in the CK group or were even significantly lower than those in the CK group ( P< P < 0.05) (low and medium-dose groups of GLVO-2). At this time, the TC content in the plasma of the mice in the administration groups had returned to the normal level. After the treatment ended, GLVO-1, GLVO-2, and GLVO-3 could reduce the TC level to 72.72%, 71.56%, and 54.75% of the model group at the lowest respectively.

[0060] At Figure 7After 10 weeks of treatment, the TG levels in each drug administration group were significantly lower than those in the model group ( P< 0.05 or P< 0.01). At this time, the TG content in the plasma of the mice in the drug administration group had returned to the normal level and maintained a relatively stable state. After the treatment ended, GLVO-1, GLVO-2, and GLVO-3 could reduce the TG level to 56.58%, 54.59%, and 48.82% of the model group at the lowest respectively.

[0061] Example 5: Proliferation-promoting effect of volatile oil from Glechoma longituba on vascular endothelial cells Using a 25 cm 2 culture flask, inoculate SVEC4-10 cells into the complete culture medium containing 10% fetal bovine serum and 1% penicillin / streptomycin, and place them in an incubator at 37 °C and 5% CO2 for culture. Take out the cells from the culture medium, observe the cell growth status under an inverted microscope, and replace the complete culture medium in a timely manner according to the cell growth status and the degree of yellowing of the culture medium color. When changing the medium, aspirate the original culture medium in the culture flask, wash the surface 3 times with PBS, add the new complete culture medium, and put it back into the incubator at 37 °C and 5% CO2 for continued culture. When the cell growth reaches a coverage rate of 80%, aspirate the original culture medium, wash 3 times with PBS, add 2 mL of 0.25% trypsin to digest for 5 min, observe under the microscope that the cells become round and detached, and quickly transfer them to the ultra-clean workbench and add 6 mL of complete culture medium to terminate the digestion. Gently pipette the cells to make them completely detached and then aspirate them into a centrifuge tube, centrifuge (1000 r / min, 5 min) and discard the supernatant, add the new complete culture medium to resuspend the cells, passage them at a ratio of 1:5, and take the cells in the logarithmic growth phase for the experiment.

[0062] Take the cells in the logarithmic growth phase (the cell coverage rate in the culture flask is 70% to 90%), digest them with 0.25% trypsin for 5 min to make the cells detached, terminate the digestion and collect the cells, centrifuge at 1000 r / min for 5 min, then aspirate the supernatant, and resuspend them with the complete medium to make a cell suspension of 1.5 × 10 4 cells / mL. Inoculate the cells into a 96-well plate, add 0.2 mL of cell suspension to each well (the volume of each medium change is 0.2 mL), culture for 48 h to make the cell coverage rate reach 60%. Aspirate the original medium in the wells and replace it with serum-free medium, culture for 24 h to make the cell coverage rate reach 70%.

[0063] Divide the cells into a blank group, a control group, and a drug administration group. The blank group is the complete culture medium without cells and containing the solvent; the control group is treated with the complete culture medium containing the solvent; the drug administration group is respectively added with the complete culture medium containing different concentrations of volatile oil from Glechoma longituba (GLVO). After culturing for 24 h, use CCK-8 to measure the cell viability. The formula for measuring cell viability is as follows: Cell viability (%) = (A experimental group - A blank group) / (A control group - A blank group) × 100 In Figure 8 , GLVO-1, GLVO-2, GLVO-3, GLVO-4, and GLVO-5 represent the essential oils of the control group, 1.6 wt / v% stressed for 4 h (essential oil-1), 0.4 wt / v% stressed for 72 h (essential oil-2), 1.6 wt / v% stressed for 96 h (essential oil-3), and 1.6 wt / v% stressed for 48 h (essential oil-4), respectively. The essential oil of Glechoma longituba (Nakai) Kupr. can significantly improve the viability of SVEC4-10 cells. The promoting effects of GLVO-1, GLVO-3, and GLVO-5 are more obvious and show a dose-dependence in the concentration range of 1.5625 - 6.25 μg / mL. Under the action of the three essential oils, the viability of SVEC4-10 cells reaches the maximum value at a concentration of 6.25 μg / mL, increasing to (295.63 ± 14.08)%, (283.27 ± 8.99)%, and (173.02 ± 37.27)%, respectively. GLVO-2 and GLVO-4 have a significant promoting effect on the viability of SVEC4-10 cells and show a dose-dependence in the concentration range of 3.125 - 12.5 μg / mL. The cell viability reaches the maximum value at a concentration of 12.5 μg / mL for both, being (312.92 ± 14.33)% and (185.89 ± 35.97)%, respectively.

[0064] Example 6: Repair effect of the essential oil of Glechoma longituba (Nakai) Kupr. on vascular endothelial cells damaged by LPS Cells in the logarithmic growth phase were digested with trypsin and prepared into a cell suspension of 10 6 cells / mL, inoculated into 12-well plates, 2 mL per well, and cultured for 48 h. When the cell confluence reached 70%, the supernatant was aspirated and replaced with serum-free medium, and cultured for 24 h. When the cells grew to 80%, the supernatant was aspirated, and complete medium containing 1.5 μg / mL LPS was added. A control group was set. After culturing for 24 h, the damaged cells were divided into a model group, a positive group, and a drug administration group. The supernatant of all cells was aspirated, and different complete culture media were added. The control group and the model group contained only the solvent, the positive group contained 100 μg / mL of VE, and the drug administration group contained different concentrations of GLVO, and cultured for 24 h. The cell viability was measured using a CCK-8 kit.

[0065] The cells were lysed with RIPA lysis buffer and centrifuged (14000 r / min, 5 min), and the supernatant was taken. The total protein and MDA contents of the supernatant were measured according to the kit instructions.

[0066] In Figure 9Among them, the cell viability of the low, medium, and high-dose treatment groups of GLVO and the positive group (VE) was significantly increased ( P< P < 0.01). Among them, the high-dose groups of GLVO-1 and GLVO-4 were significantly higher than the CK group ( P< P < 0.05). The low, medium, and high-dose groups of GLVO-2, GLVO-3, and GLVO-5 were all higher than the CK group ( P< P < 0.05). GLVO-1, GLVO-2, GLVO-3, GLVO-4, and GLVO-5 could increase the cell viability to (132.82 ± 7.57)%, (167.56 ± 5.64)%, (153.96 ± 7.70)%, (126.82 ± 4.72)%, and (137.15 ± 8.20)%, respectively.

[0067] In Figure 10 , the MDA content level of the model group was higher than that of other groups, which was (1.43 ± 0.25) nmol / mgprot. After GLVO treatment for 24 h, the MDA level in SVEC4-10 cells could be reduced in a dose-dependent manner. The low, medium, and high-dose groups of GLVO-5 were all lower than the model group and the blank group, and the effect was better than that of GLVO-1. The MDA levels of the low, medium, and high-dose groups of GLVO-4 were all lower than those of the model group, CK group, and vitamin E group at the same time, and the lowest reduction compared with the model group was 92.05%. The reduction amplitude of the intracellular MDA level treated with GLVO-4 was the largest.

Claims

1. A Glechoma longituba essential oil, characterized in that, The volatile oil components of Glechoma longituba are obtained by extracting Glechoma longituba under Na2SO4 stress, and the method comprises the following steps: Step 1: Transfer the aseptic tissue culture seedlings of Glechoma longituba into 1 / 2 Japanese Horticultural Standard Solution for culture; Step 2: Cut the Glechoma longituba seedlings into small segments, insert them into 1 / 2 Japanese Horticultural Standard Solution for culture, the culture temperature is 20-30 °C, the culture time is 25-60 d, and culture until there are more than 15 nodes at the stem tip to obtain hydroponic Glechoma longituba; Step 3: After subjecting the hydroponic Glechoma longituba to 1.2 wt / v%-2.0 wt / v% Na2SO4 stress for 2-6 h, extract the volatile oil by steam distillation.

2. The volatile oil of Glechoma longituba according to claim 1, characterized in that, Among the top 50 compounds by content, compared with the essential oil of Glechoma longituba under non-stress conditions, 15 new compounds were added, namely 1,3,4-trimethyl-3-cyclohexene-1-carboxaldehyde (CAS No.: 040702-26-9), Z,Z,Z-1,5,9,9-tetramethyl-1,4,7-cycloundecatriene (CAS No.: 1000062-61-9), β-ocimene (CAS No.: 013877-91-3), hexahydro-2,5-methano-1H-indene-7(4H)-one (CAS No.: 027567-85-7), ethyl decanoate (CAS No.: 000110-38-3), nonenal (000124-19-6), 2-methyl-3-phenylpropenal (CAS No.: 000101-39-3), 2-methyloctane (CAS No.: 003221-61-2), trans-2-methyl-5-(1-methylethenyl)cyclohexanone (CAS No.: 005948-04-9), 1-vinyl-1-methyl-2,4-bis(1-methylethenyl)cyclohexane (CAS No.: 110823-68-2), 7,11-dimethyl-3-methylene-1,6,10-dodecatriene (CAS No.: 077129-48-7), (1-α,2-β,4-β)-1-vinyl-1-methyl-2,4-bis(1-methylethenyl)cyclohexene (CAS No.: 033880-83-0), (Z,Z)-α-farnesene (1000293-03-1), 2-methyl-5-(1-methylethyl)phenol (000499-75-2), cis-p-mentha-2,8-dien-1-ol (003886-78-0); The contents of the following 6 compounds increased significantly (P < 0.05): caryophyllene (CAS No.: 000087-44-5), (E)-β-farnesene (CAS No.: 018794-84-8), [S-(R,S -3-(1,5-dimethyl-4-hexenyl)-6-methylenecyclohexene (CAS No.: 020307-83-9), 1-(1,5-dimethyl-4-hexenyl)-4-methylbenzene (CAS No.: 000644-30-4), [S-(R,S )]-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS No.: 000495-60-3), (1S-cis)-1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-naphthalene (CAS No.: 000483-76-1).

3. A volatile oil of Glechoma longituba, characterized in that, The volatile oil components of Glechoma longituba are obtained by extracting Glechoma longituba under Na2SO4 stress, and the method comprises the following steps: Step 1: Transfer the aseptic tissue culture seedlings of Glechoma longituba into 1 / 2 Japanese Horticultural Standard Solution for culture; Step 2: Cut the Glechoma longituba seedlings into small segments, insert them into 1 / 2 Japanese Horticultural Standard Solution for culture, the culture temperature is 20-30 °C, the culture time is 25-60 d, and culture until there are more than 15 nodes at the stem tip to obtain hydroponic Glechoma longituba; Step 3: After subjecting the hydroponic Glechoma longituba to 0.2 wt / v%-0.6 wt / v% Na2SO4 stress for 68 - 76 h, extract the volatile oil by steam distillation.

4. The volatile oil of Glechoma longituba according to claim 3, characterized in that, Among the top 50 compounds in terms of content, compared with the essential oil of Glechoma longituba under non-stress conditions, 10 new compounds were added, namely cis-p-mentha-2,8-dien-1-ol (CAS No.: 003886-78-0), β-pinene (CAS No.: 000123-35-3), β-ocimene (CAS No.: 013877-91-3), ethyl decanoate (CAS No.: 000110-38-3), 1,7,7-trimethyl-bicyclo[2.2.1]hept-2-ene (CAS No.: 000464-17-5), (Z,Z)-α-farnesene (CAS No.: 1000293-03-1), (6R,7aR)-3,6-dimethyl-5,6,7,7a-tetrahydrobenzofuran-2(4H)-one (CAS No.: 038049-04-6), 1-ethoxy-4-ethylbenzene (CAS No.: 001585-06-4), 2-methyl-5-(1-methylethyl)phenol (CAS No.: 000499-75-2), 1-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2-buten-1-one (CAS No.: 035044-68-9); There are 6 compounds with a significant increase in content (P < 0.05): linalool (CAS No.: 000078-70-6), 3-methyl-6-(1-methylethenyl)-cyclohexen-1-one (CAS No.: 000491-09-8), 10,10-dimethyl-2,6-dienylbicyclo[7.2.0]undecane (CAS No.: 357414-37-0), [S-(R,S )]-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS No.: 000495-60-3), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS No.: 007216-56-0), (1S-cis)-1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-naphthalene (CAS No.: 000483-76-1).

5. A volatile oil of Glechoma longituba, characterized in that, The volatile oil components of Glechoma longituba are obtained by extracting Glechoma longituba under Na2SO4 stress, and the method comprises the following steps: Step 1: Transfer the aseptic tissue culture seedlings of Glechoma longituba into 1 / 2 Japanese Horticultural Standard Solution for culture; Step 2: Cut the Glechoma longituba seedlings into small segments, insert them into 1 / 2 Japanese Horticultural Standard Solution for culture, the culture temperature is 20-30 °C, the culture time is 25-60 d, and culture until there are more than 15 nodes at the stem tip to obtain hydroponic Glechoma longituba; Step 3: After subjecting the hydroponic Glechoma longituba to 1.2 wt / v%-2.0 wt / v% Na2SO4 stress for 92 - 100 h, extract the volatile oil by steam distillation.

6. The volatile oil of Glechoma longituba according to claim 5, characterized in that, Among the top 50 compounds in terms of content, compared with the essential oil of Glechoma longituba under non-stress conditions, 11 new compounds were added, namely: (1S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene (CAS No.: 007785-26-4), β-ocimene (CAS No.: 013877-91-3), ethyl decanoate (CAS No.: 000110-38-3), 2-methyl-3-phenyl-2-propenal (CAS No.: 000101-39-3), aromadendrene (CAS No.: 000489-39-4), α-terpineol (CAS No.: 000098-55-5), cis-3-hexenyl cis-3-hexenoate (CAS No.: 061444-38-0), (Z)-2-(3,3-dimethylcyclohexenyl)-ethanol (CAS No.: 026532-23-0), neodihydrocarveol (CAS No.: 018675-34-8), 1-methyl-3-(1-methylethenyl)cyclohexene (CAS No.: 000499-03-6), trans-2-methyl-5-(1-methylethenyl)cyclohexanone (CAS No.: 005948-04-9); the contents of the following 4 compounds were significantly increased (P < 0.05): D-limonene (CAS No.: 005989-27-5), trans-5-methyl-2-(1-methylethenyl)-cyclohexanone (CAS No.: 029606-79-9), caryophyllene (CAS No.: 000087-44-5), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS No.: 007216-56-0).

7. A volatile oil of Glechoma longituba, characterized in that, The components of the essential oil of Glechoma longituba were obtained by extracting Glechoma longituba stressed by Na2SO4, including the following steps: Step 1: Transfer the aseptic tissue culture seedlings of Glechoma longituba into 1 / 2 Japanese Horticultural Test Commodity Nutrient Solution for cultivation; Step 2: Cut the Glechoma longituba seedlings into small segments and insert them into 1 / 2 Japanese Horticultural Test Commodity Nutrient Solution for cultivation. The cultivation temperature is 20-30 °C, the cultivation time is 25-60 d, and cultivate until there are more than 15 nodes at the stem tip to obtain hydroponic Glechoma longituba; Step 3: After stressing the hydroponic Glechoma longituba with 1.2 wt / v%-2.0 wt / v% Na2SO4 for 44 - 52 h, extract the essential oil by steam distillation method.

8. The volatile oil of Glechoma longituba (Nakai) Kupr. according to claim 7, wherein, Among the top 50 compounds in terms of content, compared with the essential oil of Glechoma longituba under non-stress conditions, 11 new compounds were added, namely: β-ocimene (CAS No.: 013877-91-3), ethyl caprate (CAS No.: 000110-38-3), Z,Z,Z-1,5,9,9-tetramethyl-1,4,7-cycloundecatriene (CAS No.: 1000062-61-9), hexahydro-2,5-methano-1H-indene-7(4H)-one (CAS No.: 027567-85-7), 2-isopropyl-5-methyl-3-cyclohexen-1-one (CAS No.: 1000155-47-0), (+)-4-carene (CAS No.: 029050-33-7), 3-methyl-6-(1-methylethenyl)-cyclohexene (CAS No.: 000586-63-0), 7,11-dimethyl-3-methylene-1,6,10-dodecatriene (CAS No.: 077129-48-7), 1,5,5-trimethyl-6-methylene-cyclohexene (000514-95-4), 5-methyl-3-(1-methylethenyl)cyclohexene (CAS No.: 056816-08-1), cis-3-hexenyl cis-3-hexenoate (CAS No.: 061444-38-0). The contents of the following 5 compounds were significantly increased (P < 0.05): trans-5-methyl-2-(1-methylethenyl)-cyclohexanone (CAS No.: 029606-79-9), myrcene (CAS No.: 000087-44-5), linalool (CAS No.: 000078-70-6), [S-(R,S )]-5-(1,5-dimethyl-4-hexenyl)-2-methyl-1,3-cyclohexadiene (CAS No.: 000495-60-3), (E,Z)-2,6-dimethyl-2,4,6-octatriene (CAS No.: 007216-56-0).

9. The essential oil of Glechoma longituba according to claims 1-8 can be used in the preparation of drugs for cardiovascular and cerebrovascular system-related diseases such as antioxidant, lipid-lowering, protecting vascular endothelial cells and anti-atherosclerosis.